Prolactin/PRL Antibody - #DF6506

Figure 1 Inhibition of miR-29a attenuated proliferation and decidualization of the human ESCs (A) The expression status of miR-29a in different stages of endometrial tissues and decidual tissues. qRT-PCR analysis of miR-29a on endometrial tissues and decidual tissues. All results were relative to endometrial tissues of proliferative phase, and U6 served as an internal control. (B) Immunofluorescence for Vimentin in human ESCs showing cellular localization. Scale bar: 50 μm. (C) Phase contrast images of human ESCs. Scale bar: 100 μm. (D) qRT-PCR analysis of miR-29a levels in decidualized ESCs transfected with Control/miR-29a antagomir relative to control. U6 served as an internal control. (E–G) (E) CCK-8 assays were performed to evaluate decidualized ESCs viability transfected with Control/miR-29a antagomirfor 48h. Western blot analysis (F) and quantification (G) of PRL and IGFBP1 in decidualized ESCs transfected with Control/miR-29a antagomir (n = 3). Data are represented as mean ± SEM. An unpaired two-tailed Student's t-test was used for conducting direct comparison between two groups. Student's t test: n.s. = not significant, ∗p < 0.05, ∗∗p < 0.01.

FIGURE 3 Effects of TBMA, hordenine, and N-methyltyramine on PRL expression. (A,B) PRL protein expression determined by western blotting using GAPDH as a loading control. Data are presented as the mean ± SD (n = 3 rats per group). (C) PRL concentrations detected by ELIA (n = 8). (D) PRL mRNA expression. mRNA expression was determined using RT-PCR (n = 3 rats per group; # p < 0.05 and ## p < 0.01 versus the control group; * p < 0.05 and ** p < 0.01 versus the Ris group. Bro = bromocriptine; Hor = hordenine; N-Methy = N-methyltyramine; Ris = risperidone; TBMA = total barley maiya alkaloids.

Figure 5. Function of 15-PGDH. (A) PGE2 levels in cell lysates were quantified by ELISA. The results are representative of at least three independent experiments. (B,C) F-actin staining revealed cytoskeletal changes in dhESCs and HTR8 cells in the NC (normal control) group and SW033291 group. (D,E) Western blot assays show the relative levels of EMT markers and HLA-G in Jeg3. (F,G) Western blot assays show the relative levels of EMT markers and decidualization markers in hESC. Each experiment was independently performed three times. CTB: cytotrophoblast; EVT: extravillous trophoblasts; EMT: epithelial–mesenchymal transition; DSC: decidual stromal cell. The experiment was repeated 3 times independently. Band intensities were quantified and normalized to the GAPDH values. Values are the mean ± SD.

Fig. 3. Effects of TBMA on PRL in MMQ cells and GH3 cells. (A) Effects of bromocriptine on PRL protein expression in MMQ cells and GH3 cells (n = 3). (B, C) Effects of TBMA on PRL in the supernatant of MMQ cells and GH3 cells detected by ELISA (n = 6). (D, E) Effects of TBMA on PRL protein in MMQ cells and GH3 cells detected by Western Blot (n = 3). Data were expressed as mean ± SD; compared with control group, **P < 0.01, *P < 0.05. TBMA = total barley maiya alkaloids; Bro = bromocriptine.

Figure 2 HI-TOPK-032 reduced PRL production in pituitary tumor cells and it worked by mediating p38 MAPK signaling pathway. (A) HI-TOPK-032 decreased PRL protein expression in pituitary tumor cells detected by Western Blot assay. (B) HI-TOPK-032 decreased PRL mRNA expression in pituitary tumor cells detected by qRT-PCR assay. (C) HI-TOPK-032 decreased PRL secretion in cell supernatant detected by Elisa assay. (D) The expressions of TOPK, p38 MAPK and its phosphorylation level in pituitary tumor cells after HI-TOPK-032 intervention were detected by Western Blot assay. (E) HI-TOPK-032 reduced the phosphorylation of TOPK. (F) HI-TOPK-032 reduced the phosphorylation of p38 MAPK. *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001.